不同表面消毒方法对核桃叶内生菌分离效果的比较

摘 要：【研究目的】表面消毒是植物内生菌分离的关键步骤之一，不同的表面消毒方法不仅会影响表面消毒效果，对内生菌的分离效果也有一定影响。本文分析比较了几种不同的表面消毒方法下分离获得的内生菌数量及种类。【方法】采用传统的分离方法，即真菌用组织块法，细菌用稀释梯度法进行分离。【结果】结果表明无论是内生真菌还是内生细菌二氧化氯方法处理下获得的内生菌种类最多。即二氧化氯法获真菌6株，鉴定为5个属，细菌37株，鉴定为11个属种（其中包括1个放线菌属）；次氯酸钠法获真菌8株，鉴定为3个属，细菌4株，鉴定为3个属种；升汞法获真菌3株，鉴定为3个属，细菌1株，为未知种。从分离内生菌的数量上比较，二氧化氯方法分离获得的细菌数显著性的多于其他两种方法，由多到少依次为二氧化氯法、次氯酸钠法、升汞法。【结论】二氧化氯表面消毒更适合进行内生菌多样性研究，但不同的表面消毒方法会获得不同的机会菌株。

Comparison of different surface disinfection methods for the isolation of the endophytes in walnut leaves

The purpose of this article was to compare the effects of disinfection on the species and quantity of the endophytes isolated from walnut leaves. The fungi were separated by the tissue isolation. The bacteria were separated by the dilution method. Then the fungi were identified based on morphological characteristics. The bacteria were identified based on the morphological characteristics and 16S rDNA sequence. The results indicated that the immersing 30 s in 0.3% ClO2 was the best way for isolating endophytes from walnut leaves, 6 endofungi identified 5 genus and 37 endophytic bacteria identified 11 species （including one actinobacteria） were isolated by this way. 8 endofugi identified 3 genus and 4 endophytic bacteria identified 3 species bacteria were isolated by NaC10, 3 endofungi identified 3 genera and 1 endophytic bacteria identified 1 specie were isolated by HgCl. The endophytic bacteria which were isolated based on the ClO2 were more than other two ways significantly. So the ClO2 should be a best method to compare the diversity of endophytes in different plants. This study also indicated that different disinfectants would isolate different endophytic groups.